Lucchinetti et al., Supplementary Material

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Lucchinetti et al., Supplementary Material

Supplementary material to:

Gene Regulatory Control of Myocardial Energy Metabolism Predicts Postoperative Cardiac Function in Patients Undergoing Off-pump Coronary Artery Bypass Graft Surgery: Role of Anesthetic Choice

by E. Lucchinetti, C. Hofer, L. Bestmann, M. Hersberger, J. Feng, M. Zhu, L. Furrer, M.C. Schaub, R. Tavakoli, M. Genoni, A. Zollinger, and M. Zaugg

including:

• Supplementary Information

• Three supplementary tables (Table S1 to Table S3)

• Three supplementary figures (Figure S1 to Figure S3)

Supplementary information

Gene Set Enrichment Analysis (GSEA) is designed to identify genes with co-ordinate transcriptional regulation within functionally related groups of genes, which are called gene sets in GSEA. Signaling pathways are examples of gene sets. Pathway information was curated using knowledge from online databases and from peer- reviewed literature sources. For further details about gene sets and statistical procedures, please see the online supplementary material to the original publication (Subramanian et al. PNAS 2005;102:15545-50), which is freely available under:

http://www.pnas.org/cgi/data/0506580102/DC2/1

Public online pathway sources are: Biocarta (http://www.biocarta.com), Human Protein Reference Database (www.hprd.org), Gene Ontology (www.geneontology.org), Signal Transduction Knowledge Environment STKE: (http://stke.sciencemag.org/cm/), GenMAPP (http://www.genmapp.org), Signaling PAthway Database SPAD (http://www.grt.kyushu-u.ac.jp/spad/menu.html), Kyoto Encyclopedia of Genes and Genomes (http://www.genome.jp/kegg/kegg2.html), the AfCS-Nature Signaling Gateway (www.signaling-gateway.org), the Cancer Genome Anatomy Project (http://cgap.nci.nih.gov/Pathways). The G-CSF pathway was curated by the authors according to the following peer-reviewed publications:

1. Harada et al. G-CSF prevents cardiac remodeling after myocardial infarction by

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2 activating the Jak-Stat pathway in cardiomyocytes. Nat Med 2005; 11: 305-11 2. Takano et al. Effects of G-CSF on left ventricular remodeling and heart failure

after acute myocardial infarction. J Mol Med 2006; 84: 185-93 GSEA was selected to achieve the following analysis goals:

Goal 1: to identify pathways that are altered from time T1 (baseline; before off-pump CABG surgery) to time T2 (end of CABG surgery) in both the SEVO and the PROP groups. This resulted in the list of pathways given in Table 4. These pathways appear to be mainly regulated by the surgical stimulus per se with no detectable modulation by the two anesthetics used.

Goal 2: to identify pathways that are differentially altered between SEVO and PROP from time T1 to time T2, which would illustrate the differential transcriptional responses to SEVO and PROP.

GSEA was first used to screen for regulated pathways using a four-phenotype design,

namely (SEVO, T1), (SEVO, T2), (PROP, T1), and (PROP, T2) as illustrated in panel A

of the following scheme:

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3 Of note, all patients received exactly the same anesthetic regimen up to the time of the first biopsy (T1). Thus, the first biopsies represent the background baseline gene expression (phenotypes (SEVO, T1) and (PROP, T1)), while the second biopsies predominantly mirror the transcriptional changes to cardiac surgery modulated by the two anesthetics (phenotypes (SEVO, T2) and (PROP, T2)). The comparison (SEVO, T1) vs. (PROP, T1) of this GSEA analysis revealed an interesting heterogeneous pattern in energy metabolism-related gene activity, reflecting the population variation in patients prior to treatment with a subtle but significant difference between the study groups. This difference in baseline gene activity occurred despite proper randomization and accurate selection of patients (diabetic patients were excluded). Therefore, the activity of the peroxisome proliferator receptor gamma co-activator 1-alpha (PGC-1α) pathway was included as one of the relevant variables in our subsequent multivariate analysis. Moreover, this first GSEA analysis also provided a global picture of the pathways regulated during off-pump CABG surgery (comparison (SEVO, T2) vs.

(SEVO, T1) and comparison (PROP, T2) vs. (PROP, T1)).

Because of the detected differences in baseline gene activity, which would have clearly biased the results, we did not compare directly SEVO vs. PROP ((SEVO, T2) vs.

(PROP, T2)) in this first GSEA analysis. In order to correct for background activity, fold

changes were computed, and GSEA analysis was again applied, this time with a two-

phenotype design, namely SEVO and PROP (please see panel B in the above

scheme). This analysis revealed the reported differentially regulated pathways of fatty

acid metabolism, DNA-damage (both enriched in PROP), and G-CSF/Jak2/STAT3

survival (the only pathway significantly enriched in SEVO).

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Affymetrix ID representative

public ID gene title forward primer reverse primer

203973_s_at NM_005195 CCAAT/enhancer binding protein

(C/EBP), delta 5' - TGC AGT TTG GGA CAT AG - 3' 5' - CTT AGC TGC ATC AAC AGG AG - 3' 206236_at NM_005282 G protein-coupled receptor 4 5' - ACT CAC CTC CAA GAG GAA CA - 3' 5' - AGA CCA GGA GAG AAG GGA CT- 3' 234103_at AU145191 Sodium- and chloride-activated ATP-

sensitive potassium channel SLICK 5' - CAG AAC AAT GGC AGA AGA TG - 3' 5' - CCC TCC TAA CAC CAA TCA AG - 3' 202484_s_at AF072242 methyl-CpG binding domain protein 2 5' -AAC AAC CGG TAA CCA AAG TC - 3' 5' - ATT GCT ACC TGG ACC AAC TC- 3' 202458_at NM_007173 protease, serine, 23 5' - CCA GAG AAC AAT GGA AGC A - 3' 5' - ATA GCT CCC AGA AAG CAC AC - 3' 203619_s_at NM_012306 Fas apoptotic inhibitory molecule 2 5' - GAA GCC TAG CAG GAT TTC AG- 3' 5' - ACC TGG ATG AGA AAG ACA GG - 3' 210501_x_at AF119846 eukaryotic translation initiation factor

3, subunit 12 5' - CTA AAG GTG TGG ATG AGC AA - 3' 5' - CCT GAA GTT ACT GGG AGG AG - 3' 243582_at AW082633 SH3 domain containing ring finger 2 5' - GAC GTT ATC CAC CTC CTC TG - 3' 5' -TTA GGA CAA GGG TCT CTT CG - 3' n.a. NG_002801 18S ribosomal RNA 5' - TGG TTG CAA AGC TGA AAC TTA

AAG - 3' 5' - AGT CAA ATT AAG CCG CAG GC - 3'

Table S1: Primers used for RT-PCR.

Lucchinetti et al., Supplementary Material

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Lucchinetti et al., Supplementary Material

Table S2: Transcript validation in randomly selected patients. Three patients are from the propofol anesthesia group (H02_P, H04_P, and H12_P) and three are from the sevoflurane anesthesia group (H11_S, H15_S, and H20_S).

CCAAT/enhancer binding protein (C/EBP), delta (NM_005195)

patient drug fold change chip fold change PCR validation status

H02_P propofol 3.91 12.5 +

H04_P propofol 5.47 5.67 +

H11_S sevoflurane 7.91 13.3 +

H12_P propofol 5.20 3.49 +

H15_S sevoflurane 6.67 22.2 + H20_S sevoflurane 5.44 6.62 + G protein-coupled receptor 4 (NM_005282)

H02_P propofol 1.76 2.64 +

H04_P propofol 1.79 1.90 +

H11_S sevoflurane 4.70 6.82 +

H12_P propofol 3.18 3.12 +

H15_S sevoflurane 3.10 2.92 + H20_S sevoflurane 2.93 5.49 + Sodium- and chloride-activated ATP-sensitive potassium channel SLICK (AU145191)

H02_P propofol 0.14 0.05 +

H04_P propofol 0.38 1.40 -

H11_S sevoflurane 0.17 0.42 +

H12_P propofol 0.12 0.07 +

H15_S sevoflurane 0.15 0.15 + H20_S sevoflurane 0.23 0.16 + methyl-CpG binding domain protein 2 (AF072242)

H02_P propofol 0.60 0.80 +

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6 H04_P propofol 0.73 0.78 +

H11_S sevoflurane 0.78 0.72 +

H12_P propofol 0.61 0.26 +

H15_S sevoflurane 0.65 0.55 + H20_S sevoflurane 0.76 0.61 + serine protease 23 (NM_007173)

H02_P propofol 0.43 0.24 +

H04_P propofol 1.25 1.16 +

H11_S sevoflurane 0.58 0.35 +

H12_P propofol 0.57 0.19 +

H15_S sevoflurane 0.24 0.22 + H20_S sevoflurane 0.76 0.50 + Fas apoptotic inhibitory molecule 2 (NM_012306)

H02_P propofol 0.87 0.99 +

H04_P propofol 2.53 2.14 +

H11_S sevoflurane 0.42 0.40 +

H12_P propofol 0.61 0.12 +

H15_S sevoflurane 1.20 0.83 - H20_S sevoflurane 0.56 0.31 + eukaryotic translation initiation factor 3, subunit 12 (AF119846)

H02_P propofol 1.02 1.14 +

H04_P propofol 0.71 0.77 +

H11_S sevoflurane 1.04 0.90 -

H12_P propofol 0.90 0.40 +

H15_S sevoflurane 1.12 1.16 +

H20_S sevoflurane 0.97 0.90 +

SH3 domain containing ring finger 2 (AW082633)

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7 H02_P propofol 0.81 0.68 +

H04_P propofol 0.47 0.88 +

H11_S sevoflurane 0.53 1.10 -

H12_P propofol 0.70 0.18 +

H15_S sevoflurane 0.94 1.34 -

H20_S sevoflurane 0.83 0.97 +

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8 Table S3: List of transcripts in the PGC-1α pathway and in the three pathways

differentially regulated by the anesthetics sevoflurane and propofol (fatty

acid oxidation pathway, G-CSF survival pathway, and DNA-damage

signaling pathway).

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PGC-1alpha

Affy probe Pgc1a_description gene symbol enriched

1487_at estrogen-related receptor alpha ERR alpha Yes

202455_at histone deacetylase 5 HDAC5 Yes

203004_s_at MADS box transcription enhancer factor 2, polypeptide D (myocyte enhancer factor 2D) MEF 2D Yes 207000_s_at protein phosphatase 3 (formerly 2B), catalytic subunit, gamma isoform (calcineurin A gamma) PPP3CC Yes 209926_at MADS box transcription enhancer factor 2, polypeptide B (myocyte enhancer factor 2B) MEF 2B Yes 210404_x_at calcium/calmodulin-dependent protein kinase (CaM kinase) II beta CaMK II beta Yes

210771_at peroxisome proliferative activated receptor, alpha PPARalpha Yes

211483_x_at calcium/calmodulin-dependent protein kinase (CaM kinase) II beta CaMK II beta Yes 212535_at MADS box transcription enhancer factor 2, polypeptide A (myocyte enhancer factor 2A) MEF 2A Yes 212669_at calcium/calmodulin-dependent protein kinase (CaM kinase) II gamma CaMK II gamma Yes 213276_at calcium/calmodulin-dependent protein kinase (CaM kinase) II beta CaMK II beta Yes 214684_at MADS box transcription enhancer factor 2, polypeptide A (myocyte enhancer factor 2A) MEF 2A Yes 219195_at peroxisome proliferative activated receptor, gamma, coactivator 1, alpha PGC-1alpha Yes

223438_s_at peroxisome proliferative activated receptor, alpha PPARalpha Yes

32540_at protein phosphatase 3 (formerly 2B), catalytic subunit, gamma isoform (calcineurin A gamma) PPP3CC Yes 32541_at protein phosphatase 3 (formerly 2B), catalytic subunit, gamma isoform (calcineurin A gamma) PPP3CC Yes

204652_s_at nuclear respiratory factor 1 NRF1 Yes

211279_at nuclear respiratory factor 1 NRF1 Yes

200622_x_at calmodulin 3 (phosphorylase kinase, delta) CALM3 No

200623_s_at calmodulin 3 (phosphorylase kinase, delta) CALM3 No

201020_at tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, eta polypeptide YWHAH No 202425_x_at protein phosphatase 3 (formerly 2B), catalytic subunit, alpha isoform (calcineurin A alpha) PPP3CA No 202429_s_at protein phosphatase 3 (formerly 2B), catalytic subunit, alpha isoform (calcineurin A alpha) PPP3CA No 202432_at protein phosphatase 3 (formerly 2B), catalytic subunit, beta isoform (calcineurin A beta) PPP3CB No 202457_s_at protein phosphatase 3 (formerly 2B), catalytic subunit, alpha isoform (calcineurin A alpha) PPP3CA No 203003_at MADS box transcription enhancer factor 2, polypeptide D (myocyte enhancer factor 2D) MEF 2D No

203193_at estrogen-related receptor alpha ERR alpha No

204392_at calcium/calmodulin-dependent protein kinase I CAMK1 No

205124_at MADS box transcription enhancer factor 2, polypeptide B (myocyte enhancer factor 2B) MEF 2B No 206603_at solute carrier family 2 (facilitated glucose transporter), member 4 GLUT4 No

207613_s_at calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CaMK II alpha No 207968_s_at MADS box transcription enhancer factor 2, polypeptide C (myocyte enhancer factor 2C) MEF 2C No

208095_s_at signal recognition particle 72kDa SRP72 No

208328_s_at MADS box transcription enhancer factor 2, polypeptide A (myocyte enhancer factor 2A) MEF 2A No 209199_s_at MADS box transcription enhancer factor 2, polypeptide C (myocyte enhancer factor 2C) MEF 2C No 209200_at MADS box transcription enhancer factor 2, polypeptide C (myocyte enhancer factor 2C) MEF 2C No

209563_x_at calmodulin 1 (phosphorylase kinase, delta) CALM1 No

209697_at protein phosphatase 3 (formerly 2B), catalytic subunit, gamma isoform (calcineurin A gamma) PPP3CC No 209817_at protein phosphatase 3 (formerly 2B), catalytic subunit, beta isoform (calcineurin A beta) PPP3CB No 209956_s_at calcium/calmodulin-dependent protein kinase (CaM kinase) II beta CaMK II beta No

210349_at calcium/calmodulin-dependent protein kinase IV CAMK4 No

211984_at calmodulin 1 (phosphorylase kinase, delta) CALM1 No

212757_s_at calcium/calmodulin-dependent protein kinase (CaM kinase) II gamma CaMK II gamma No 213108_at calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CaMK II alpha No

213688_at calmodulin 1 (phosphorylase kinase, delta) CALM1 No

213710_s_at Calmodulin 1 (phosphorylase kinase, delta) CALM1 No

213950_s_at Protein phosphatase 3 (formerly 2B), catalytic subunit, gamma isoform (calcineurin A gamma) PPP3CC No 214322_at calcium/calmodulin-dependent protein kinase (CaM kinase) II gamma CaMK II gamma No

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PGC-1alpha

215161_at calcium/calmodulin-dependent protein kinase IG CaMKI gamma No

217128_s_at calcium/calmodulin-dependent protein kinase IG CaMKI gamma No

34846_at Calcium/calmodulin-dependent protein kinase (CaM kinase) II beta CaMK II beta No

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G-CSF_survival

Affy probe G-CSF_description gene symbol enriched

215037_s_at BCL2-like 1 BCL2L1 Yes

207082_at colony stimulating factor 1 (macrophage) CSF1 Yes

210229_s_at colony stimulating factor 2 (granulocyte-macrophage) CSF2 Yes

207442_at colony stimulating factor 3 (granulocyte) CSF3 Yes

205841_at Janus kinase 2 (a protein tyrosine kinase) JAK2 Yes

205842_s_at Janus kinase 2 (a protein tyrosine kinase) JAK2 Yes

211108_s_at Janus kinase 3 (a protein tyrosine kinase, leukocyte) JAK3 Yes

202743_at phosphoinositide-3-kinase, regulatory subunit 3 (p55, gamma) PIK3R3 Yes 211580_s_at phosphoinositide-3-kinase, regulatory subunit 3 (p55, gamma) PIK3R3 Yes 208991_at signal transducer and activator of transcription 3 (acute-phase response factor) STAT3 Yes 208992_s_at signal transducer and activator of transcription 3 (acute-phase response factor) STAT3 Yes

203808_at v-akt murine thymoma viral oncogene homolog 2 AKT2 Yes

212171_x_at vascular endothelial growth factor VEGF Yes

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FA_oxidation

D

Affy probe FAO_description gene symbol enriched

218322_s_at acyl-CoA synthetase long-chain family member 5 ACSL5 Yes

211207_s_at acyl-CoA synthetase long-chain family member 6 ACSL6 Yes

219986_s_at acyl-Coenzyme A dehydrogenase family, member 10 ACAD10 Yes

206068_s_at acyl-Coenzyme A dehydrogenase, long chain ACADL Yes

206069_s_at acyl-Coenzyme A dehydrogenase, long chain ACADL Yes

226030_at acyl-Coenzyme A dehydrogenase, short/branched chain ACADSB Yes

200710_at acyl-Coenzyme A dehydrogenase, very long chain ACADVL Yes

209601_at acyl-Coenzyme A oxidase 1, palmitoyl ACOX1 Yes

205364_at acyl-Coenzyme A oxidase 2, branched chain ACOX2 Yes

207820_at alcohol dehydrogenase 1A (class I), alpha polypeptide ADH1A Yes

209614_at alcohol dehydrogenase 1A (class I), alpha polypeptide /// alcohol dehydrogenase I H1C; ADH1A; ADH Yes

206262_at alcohol dehydrogenase 1C (class I), gamma polypeptide ADH1C Yes

208847_s_at alcohol dehydrogenase 5 (class III), chi polypeptide ADH5 Yes

209613_s_at alcohol dehydrogenase IB (class I), beta polypeptide ADH1B Yes

209612_s_at alcohol dehydrogenase IB (class I), beta polypeptide ADH1B Yes

212224_at aldehyde dehydrogenase 1 family, member A1 ALDH1A1 Yes

202054_s_at aldehyde dehydrogenase 3 family, member A2 ALDH3A2 Yes

207583_at ATP-binding cassette, sub-family D (ALD), member 2 ABCD2 Yes

204573_at carnitine O-octanoyltransferase CROT Yes

227468_at carnitine palmitoyltransferase 1C CPT1C Yes

204264_at carnitine palmitoyltransferase II CPT2 Yes

204263_s_at carnitine palmitoyltransferase II CPT2 Yes

241929_at CD36 antigen (collagen type I receptor, thrombospondin receptor) CD36 Yes 236923_x_at CD36 antigen (collagen type I receptor, thrombospondin receptor) CD36 Yes 242197_x_at CD36 antigen (collagen type I receptor, thrombospondin receptor) CD36 Yes

203475_at cytochrome P450, family 19, subfamily A, polypeptide 1 CYP19A1 Yes

219903_s_at cytochrome P450, family 2, subfamily C, polypeptide 8 CYP2C8 Yes

214235_at cytochrome P450, family 3, subfamily A, polypeptide 5 CYP3A5 Yes

210096_at cytochrome P450, family 4, subfamily B, polypeptide 1 CYP4B1 Yes

238032_at dehydrogenase/reductase (SDR family) member 3 DHRS3 Yes

201135_at enoyl Coenzyme A hydratase, short chain, 1, mitochondrial ECHS1 Yes

205222_at enoyl-Coenzyme A, hydratase/3-hydroxyacyl Coenzyme A dehydrogenase EHHADH Yes

243574_at L-3-hydroxyacyl-Coenzyme A dehydrogenase, short chain HADHSC Yes

201036_s_at L-3-hydroxyacyl-Coenzyme A dehydrogenase, short chain HADHSC Yes

211569_s_at L-3-hydroxyacyl-Coenzyme A dehydrogenase, short chain HADHSC Yes

239516_at Lysophospholipase-like 1 LYPLAL1 Yes

218869_at malonyl-CoA decarboxylase MLYCD Yes

226728_at solute carrier family 27 (fatty acid transporter), member 1 SLC27A1 Yes 222217_s_at solute carrier family 27 (fatty acid transporter), member 3 SLC27A3 Yes 225779_at solute carrier family 27 (fatty acid transporter), member 4 SLC27A4 Yes

214763_at thioesterase, adipose associated THEA Yes

202366_at acyl-Coenzyme A dehydrogenase, C-2 to C-3 short chain ACADS No

202502_at acyl-Coenzyme A dehydrogenase, C-4 to C-12 straight chain ACADM No

1556923_at acyl-Coenzyme A oxidase 1, palmitoyl ACOX1 No

207656_s_at acyl-Coenzyme A oxidase 1, palmitoyl ACOX1 No

243817_at acyl-Coenzyme A oxidase 3, pristanoyl ACOX3 No

204242_s_at acyl-Coenzyme A oxidase 3, pristanoyl ACOX3 No

204241_at acyl-Coenzyme A oxidase 3, pristanoyl ACOX3 No

210687_at carnitine palmitoyltransferase 1A (liver) CPT1A No

210688_s_at carnitine palmitoyltransferase 1A (liver) CPT1A No

203634_s_at carnitine palmitoyltransferase 1A (liver) CPT1A No

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FA_oxidation

203633_at carnitine palmitoyltransferase 1A (liver) CPT1A No

210070_s_at choline kinase beta /// carnitine palmitoyltransferase 1B (muscle) CHKB; CPT1B No 210069_at choline kinase beta /// carnitine palmitoyltransferase 1B (muscle) CHKB; CPT1B No

200789_at enoyl Coenzyme A hydratase 1, peroxisomal ECH1 No

208629_s_at hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-C HADHA No 208630_at hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-C HADHA No 208631_s_at hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-C HADHA No 201007_at hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-C HADHB No

201035_s_at L-3-hydroxyacyl-Coenzyme A dehydrogenase, short chain HADHSC No

210636_at peroxisome proliferative activated receptor, delta PPARD No

208044_s_at peroxisome proliferative activated receptor, delta PPARD No

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DNA_damage_signaling

Affy probe DNA_DAMAGE_SIGNALING_description gene symbol enriched

210027_s_at APEX nuclease (multifunctional DNA repair enzyme) 1 APEX1 Yes

212672_at ataxia telangiectasia mutated (includes complementation groups A, C an ATM Yes

209902_at ataxia telangiectasia and Rad3 related ATR Yes

209903_s_at ataxia telangiectasia and Rad3 related ATR Yes

208860_s_at alpha thalassemia/mental retardation syndrome X-linked (RAD54 homo ATRX Yes 211022_s_at alpha thalassemia/mental retardation syndrome X-linked (RAD54 homo ATRX Yes

211833_s_at BCL2-associated X protein BAX Yes

201848_s_at BCL2/adenovirus E1B 19kDa interacting protein 3 BNIP3 Yes

202870_s_at CDC20 cell division cycle 20 homolog (S. cerevisiae) CDC20 Yes

204696_s_at cell division cycle 25A CDC25A Yes

201853_s_at cell division cycle 25B CDC25B Yes

202246_s_at cyclin-dependent kinase 4 CDK4 Yes

205709_s_at CDP-diacylglycerol synthase (phosphatidate cytidylyltransferase) 1 CDS1 Yes

210416_s_at CHK2 checkpoint homolog (S. pombe) CHEK2 Yes

205162_at excision repair cross-complementing rodent repair deficiency, compleme CKN1 Yes 203409_at damage-specific DNA binding protein 2, 48kDa /// LIM homeobox 3 DDB2 Yes

218194_at small fragment nuclease DKFZP566E144 Yes

213468_at excision repair cross-complementing rodent repair deficiency, compleme ERCC2 Yes 202176_at excision repair cross-complementing rodent repair deficiency, compleme ERCC3 Yes 202414_at excision repair cross-complementing rodent repair deficiency, compleme ERCC5 Yes

215942_s_at G-2 and S-phase expressed 1 GTSE1 Yes

219019_at leucine-rich repeats and death domain containing LRDD Yes

203686_at N-methylpurine-DNA glycosylase MPG Yes

209421_at mutS homolog 2, colon cancer, nonpolyposis type 1 (E. coli) MSH2 Yes

213806_at purine-rich element binding protein A PURA Yes

204020_at purine-rich element binding protein A PURA Yes

211228_s_at RAD17 homolog (S. pombe) RAD17 Yes

207405_s_at RAD17 homolog (S. pombe) RAD17 Yes

210826_x_at RAD17 homolog (S. pombe) RAD17 Yes

201046_s_at RAD23 homolog A (S. cerevisiae) RAD23A Yes

208393_s_at RAD50 homolog (S. cerevisiae) RAD50 Yes

204828_at RAD9 homolog A (S. pombe) RAD9A Yes

219370_at reprimo, p53 dependent G2 arrest mediator candidate RPRM Yes

207113_s_at tumor necrosis factor (TNF superfamily, member 2) TNF Yes

204781_s_at Fas (TNF receptor superfamily, member 6) TNFRSF6 Yes

215719_x_at Fas (TNF receptor superfamily, member 6) TNFRSF6 Yes

204780_s_at Fas (TNF receptor superfamily, member 6) TNFRSF6 Yes

216252_x_at Fas (TNF receptor superfamily, member 6) TNFRSF6 Yes

201746_at tumor protein p53 (Li-Fraumeni syndrome) TP53 Yes

202330_s_at uracil-DNA glycosylase UNG Yes

209375_at xeroderma pigmentosum, complementation group C XPC Yes

203655_at X-ray repair complementing defective repair in Chinese hamster cells 1 XRCC1 Yes 207598_x_at X-ray repair complementing defective repair in Chinese hamster cells 2 XRCC2 Yes 208442_s_at ataxia telangiectasia mutated (includes complementation groups A, C an ATM No 210858_x_at ataxia telangiectasia mutated (includes complementation groups A, C an ATM No 208861_s_at alpha thalassemia/mental retardation syndrome X-linked (RAD54 homo ATRX No 208859_s_at alpha thalassemia/mental retardation syndrome X-linked (RAD54 homo ATRX No

204531_s_at breast cancer 1, early onset BRCA1 No

211851_x_at breast cancer 1, early onset BRCA1 No

204695_at cell division cycle 25A CDC25A No

205167_s_at cell division cycle 25C CDC25C No

217010_s_at cell division cycle 25C CDC25C No

203968_s_at CDC6 cell division cycle 6 homolog (S. cerevisiae) CDC6 No

203967_at CDC6 cell division cycle 6 homolog (S. cerevisiae) CDC6 No

202284_s_at cyclin-dependent kinase inhibitor 1A (p21, Cip1) CDKN1A No

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DNA_damage_signaling

205393_s_at CHK1 checkpoint homolog (S. pombe) CHEK1 No

205394_at CHK1 checkpoint homolog (S. pombe) CHEK1 No

208619_at damage-specific DNA binding protein 1, 127kDa DDB1 No

203720_s_at excision repair cross-complementing rodent repair deficiency, compleme ERCC1 No 203719_at excision repair cross-complementing rodent repair deficiency, compleme ERCC1 No 210158_at excision repair cross-complementing rodent repair deficiency, compleme ERCC4 No

204768_s_at flap structure-specific endonuclease 1 FEN1 No

204767_s_at flap structure-specific endonuclease 1 FEN1 No

204318_s_at G-2 and S-phase expressed 1 GTSE1 No

204315_s_at G-2 and S-phase expressed 1 GTSE1 No

211040_x_at G-2 and S-phase expressed 1 /// G-2 and S-phase expressed 1 GTSE1 No

204883_s_at HUS1 checkpoint homolog (S. pombe) HUS1 No

217618_x_at HUS1 checkpoint homolog (S. pombe) HUS1 No

204884_s_at HUS1 checkpoint homolog (S. pombe) HUS1 No

211832_s_at Mdm2, transformed 3T3 cell double minute 2, p53 binding protein (mous MDM2 No 217373_x_at Mdm2, transformed 3T3 cell double minute 2, p53 binding protein (mous MDM2 No 205386_s_at Mdm2, transformed 3T3 cell double minute 2, p53 binding protein (mous MDM2 No

210947_s_at mutS homolog 3 (E. coli) MSH3 No

205887_x_at mutS homolog 3 (E. coli) MSH3 No

211450_s_at mutS homolog 6 (E. coli) MSH6 No

211449_at mutS homolog 6 (E. coli) MSH6 No

202911_at mutS homolog 6 (E. coli) MSH6 No

222063_s_at CDP-diacylglycerol synthase (phosphatidate cytidylyltransferase) 1 null No

216914_at cell division cycle 25C null No

1555186_at cyclin-dependent kinase inhibitor 1A (p21, Cip1) null No

239310_at Damage-specific DNA binding protein 2, 48kDa null No

205385_at Mdm2, transformed 3T3 cell double minute 2, p53 binding protein (mous null No

204021_s_at purine-rich element binding protein A PURA No

210216_x_at RAD1 homolog (S. pombe) RAD1 No

204460_s_at RAD1 homolog (S. pombe) RAD1 No

204461_x_at RAD1 homolog (S. pombe) RAD1 No

211300_s_at tumor protein p53 (Li-Fraumeni syndrome) TP53 No

221642_at three prime repair exonuclease 1 TREX1 No

34689_at three prime repair exonuclease 1 TREX1 No

205875_s_at three prime repair exonuclease 1 TREX1 No

Zaugg_Suppl_Table3_rev.xls 2 of 2

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9 Supplementary Figure Legends

Figure S1 Heat maps of the fifty top-ranked up- and downregulated genes in propofol (PROP) patients at T1 and T2 (Suppl. Figure 1A) and sevoflurane (SEVO) patients at T1 and T2 (Suppl. Figure 1B). Blue color indicates jointly regulated transcripts between sevoflurane and propofol.

Figure S2 Heat maps of the constitutive (preoperative) gene expression of the OXPHOS pathway (Suppl. Figure 2A) and the fatty acid metabolism pathway (Suppl. Figure 2B) in the enrolled patients, marked on top by their randomization code H01 to H20 and the label S for sevoflurane or P for propofol.

Figure S3 Gene Set Enrichment Analysis (GSEA) of chip data (Affymetrix U34A) obtained from isolated perfused rat hearts exposed to isoflurane, another ether-based anesthetic gas, showing similar downregulation of genes involved in fatty acid metabolism. The rat hearts were exposed to 15 min of 2.1 vol.% isoflurane followed by 180 min of reperfusion, as previously described in detail.

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